Family of prohormone convertases in Lymnaea: Characterization of two alternatively spliced furin-like transcripts and cell-specific regulation of their expression

Sabine Spijker, August B. Smit, Hilary E. Sharp-Baker, René Van Elk, Ellen R. Van Kesteren, Jan Van Minnen, Alexander Kurosky, Wijnand P M Geraerts

    Research output: Contribution to journalArticle

    9 Citations (Scopus)

    Abstract

    The majority of neuropeptides in Lymnaea stagnalis are proteolytically processed from larger precursors at sites composed of single or multiple basic amino acid residues. Previous studies have identified several putative prohormone convertases in the brain of Lymnaea. To characterize the complete family, we undertook three independent approaches: reverse-transcribed polymerase chain reaction screening, and low-stringency cDNA and genomic library screenings. The central nervous system cDNA library screening yielded two cDNAs encoding Lfurin1 and its variant form, Lfurin1-X. Both proteins show the characteristic organization of (human) furin with a putative catalytic domain, a P domain, a Cys-rich domain, a transmembrane domain, and a cytoplasmic tail. Lfurin1 and Lfurin1-X are identical, apart from a putative alternatively spliced noncatalytic luminal protein domain, which is present exclusively in Lfurin1-X. In situ hybridization revealed that the Lfur1 gene is expressed throughout the Lymnaea brain, but that the level varies considerably from one neuron to another. Quantitative analysis of the expression level of the two alternatively spliced transcripts revealed that it is neuron type-specifically regulated. This probably indicates the functional importance of noncatalytic luminal protein domains in these enzymes. In addition, our findings suggest that apart from the identified convertases LPC2, Lfurin1/Lfurin1-X, and Lfurin2, additional prohormone convertase diversity is either not present or present only at low levels in the Lymnaea brain. Alternatively, additional prohormone convertases could exist with a lower degree of sequence conservation than the other Lymnaea prohormone convertase members. From our findings, it appears that the majority of prohormone processing in Lymnaea is carried out by the three thus far identified types of Kex2-related prohormone convertases despite the large number of neuropeptide precursors and diverse multiple basic cleavage sites hydrolyzed.

    Original languageEnglish (US)
    Pages (from-to)399-413
    Number of pages15
    JournalJournal of Neurobiology
    Volume41
    Issue number3
    DOIs
    StatePublished - Nov 15 1999

    Fingerprint

    Proprotein Convertases
    Furin
    Lymnaea
    Neuropeptides
    Gene Library
    Brain
    Neurons
    Basic Amino Acids
    Genomic Library
    In Situ Hybridization
    Tail
    Catalytic Domain
    Central Nervous System
    Complementary DNA
    Polymerase Chain Reaction
    Enzymes
    Genes

    Keywords

    • Central nervous system
    • Mollusk Lymnaea stagnalis
    • Neuropeptide biosynthesis
    • Quantitative in situ hybridization
    • Subtilisin- like prohormone convertases

    ASJC Scopus subject areas

    • Neuroscience(all)

    Cite this

    Family of prohormone convertases in Lymnaea : Characterization of two alternatively spliced furin-like transcripts and cell-specific regulation of their expression. / Spijker, Sabine; Smit, August B.; Sharp-Baker, Hilary E.; Van Elk, René; Van Kesteren, Ellen R.; Van Minnen, Jan; Kurosky, Alexander; Geraerts, Wijnand P M.

    In: Journal of Neurobiology, Vol. 41, No. 3, 15.11.1999, p. 399-413.

    Research output: Contribution to journalArticle

    Spijker, Sabine ; Smit, August B. ; Sharp-Baker, Hilary E. ; Van Elk, René ; Van Kesteren, Ellen R. ; Van Minnen, Jan ; Kurosky, Alexander ; Geraerts, Wijnand P M. / Family of prohormone convertases in Lymnaea : Characterization of two alternatively spliced furin-like transcripts and cell-specific regulation of their expression. In: Journal of Neurobiology. 1999 ; Vol. 41, No. 3. pp. 399-413.
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